载脂蛋白E基因分析中血样处理方法比较研究

Comparing studies of blood sample treatments for ApoE gene polymorphism analysis

目的为筛选快速、简便、灵敏的适用于大规模流行病学调查的载脂蛋白E(ApoE)基因分析的血样处理方法,对不同血样处理方法进行比较研究。方法建立本实验室ApoE基因分析体系,并对蒸馏水萃取法、血细胞裂解法、Tris-EDTA(TE)萃取法等血样处理方法进行了比较。结果在蒸馏水萃取法、血细胞裂解法、TE萃取法中提取效率最高的是血细胞裂解法,蒸馏水萃取法次之,TE萃取法最低;在灵敏度方面,蒸馏水萃取法灵敏度最高;在血片干燥程度对PCR结果的影响中,只有彻底干燥24h后进行处理,才能获得清晰的PCR结果;在经济方面,本实验室体系可获得与使用进口试剂盒同样的检测效果,而进口试剂盒价格昂贵。结论蒸馏水萃取法非常适合大规模流行病学调查,具备快速、简便、灵敏等优点。

Objective For the larger scale epidemiological study on apolipoprotein E(ApoE) gene polymorphism analysis.To compare the different blood sample treatments.Methods ApoE allele polymorphism analysis enthod was established in our lab,and the different trace DNA extraction methods of destilled-water extraction method,blood cell lysis method and Tris-EDTA extraction method were compared.Results The most sensitive method was distilled-water extraction method.As to the effect of blood dryness on filter,the best result could be achieved from over 24?h drying.As to economy and convenience of the methods,this laboratory's ApoE analysis system had the same results with the import reagent kit but cheaper.Conclusion The distilled-water extraction method for ApoE allele polymorphism analysis was proper for larger scale epidemiological study,which had the advantages of quick,convenient and sensitive characters.